Kahalalide F and compositions and uses thereof

ABSTRACT

KalahideKahalalide F, of formula I below, may be isolated from a sacoglossan. The compound may be used in the manufacture of pharmaceutical compositions or in the treatment of tumors or viral conditions

This invention is concerned with a cytotoxic and antiviral compoundisolated from the sacoglossan, Elysia rafescens rufescens.

According to the invention there is provided, a the new compound, thepeptide, Kahalalide F, of the formula I:

The antitumor activities of this compound has been determined “in vitro”in cell cultures of human lung carcinoma A-549 and human colon carcinomaHT-29. The procedure was carried out using the metnhodology methodologydescribed by Raymond J. Bergeron et al. Biochem. Bioph. Res. Comm. 1984,121(3), 848-854 and by Alan C. Schroeder et al. J. Med. Chem. 1981, 241078-1083.

The antiviral activities of this compound the compound Kahalalide F havealso been determined “in vitro” against HSV (Herpes simplex virus) andVSV (Vesicular stomatitis virus). The methodology used to carry out thisdetermination is described by Raymond J. Bergeron et al. Biochem. Bioph.Res. Comm. 1984, 121(3), 848-854 and by Alan C. Schroeder et al. J. Med.Chem. 1981, 24 1078-1083.

Therefore, the present invention also provides a method of treating anymammal affected by a malignant tumor sensitive to compounds abovedescribed, which comprises administering to the affected individual atherapeutically effective amount of these compounds or a pharmaceuticalcomposition thereof; and a method of treating viral infections inmammals, comprising administering to a patient in need of suchtreatment, an antiviral effective amount of the compounds described inthe present invention.

The present invention also relates to pharmaceutical preparations whichcontain as active ingredient these compounds, or a pharmaceuticallyacceptable acid addition salt thereof, as well as the process for itspreparation.

Examples of pharmaceutical compositions include any solid (tablets,pills, capsules, granules, etc.) or liquid (solutions, suspensions oremulsions) suitable composition for oral, topical or parenteraladministration, and they may contain the pure compound or in combinationwith any carrier or other pharmacologically active compounds. Thesecompositions may need to be sterile when administered parenterally.

The correct dosage of a pharmaceutical composition of these compoundsthis compound will vary according to the particular formulation, themode of application and particular site, host and tumor being treated.Other factors like age, body weight, sex, diet, time of administration,rate of excretion, condition of the host, drug combinations, reactionsensitivities and severity of disease shall be taken in account.Administration can be carried out continuously or periodically withinthe maximum tolerated dose.

Kahalalide F was isolated from the sacoglossan, Elysia rufescens Elysiarufescens (family Plakobranchidae, order Sacoglossa), collected nearBlack point Point, Oahu. This animal varies in size between 1 and 4 cm;it is dark red-brown in color with light-colored spots. There is orangefringing of the parapodia, which have very small dark green spots fromsequestered chloroplasts. Elysia rufescens feeds on the delicate,feather-like green alga Bryopsis Bryopsis sp. ¹ Kahalalide F can also beisolated from this alga. Two hundred animals were collected over theperiod of several weeks during the spring, of 1991 and extracted withEtOH. The extracts were then chromatographed by silica gel flashchromatography (hexane, hexane/EtOAc (1:1), EtOAc, EtOAc (1:1)EtOAc/MeOH ( 1:1 ), MeOH and MeOH/HOAc (98:2)). The peptides were elutedwith EtOAc/MeOH (1:1). Final purification was accomplished by repeatedHPLC (RP C18) using MeCN/H₂O with 0.1% TFA (70-45% H₂O).

The structures of the peptides were elucidated by 2D NMR experiments(HMQC, HMBC, TOCSY, COSY and ROESY).

Kahalalide F was isolated as a white amorphous powder in 0.02% 0.01%yield. A molecular formula of C₇₅H₁₂₄N₁₄O₁₆ was deduced from detailedanalyses of the ¹³C and ¹H NMR spectra and the high resolution FAB massspectrum. The 14 substructures in this compound arise from five valines,two isoleucines, two threonines, ornithine, dehydroaminobutyric acid. ,proline, phenilalanine phenylalanine and 5-methythexanoic5-methylhexanoic acid (5-MeHex). Kahalalide F is the largest peptide inthis series of compounds.

EXPERIMENTAL

General Considerations

Optical rotations were measured on a Jasco DIP-370 digital polarimeter.Infrared spectra were recorded on a Nicolet MX-5 FTIR spectrometer. Gaschromatography was accomplished using a Hewlett-Packard Model 5890instrument. Mass spectra were measured on a VG-70SE magnetic sector massspectrometer. NMR spectra were measured on a General Electric QE-300 ora GN OMEGA 500 instrument. ¹H NMR chemical shifts are reported in ppmwith the chemical shift of the residual protons of the solvent used asinternal standards. ¹³C NMR chemical shifts are reported in ppm by usingthe natural abundance ¹³C of the solvent as an internal standard.Ultraviolet spectra were recorded on a Hewlett-Packard Model 8452A diodearray spectrophotometer. All solvents were distilled from glass beforeuse.

Two hundred sacoglossans (Elysia rufescens), were collected at BlackPoint, O'ahu during April and May 1992 1991, and extracted 3 times withEtOH. Spring appears to be the time of year Elysia rufescens is ingreatest abundance at Black Point. The combined extracts were thenchromatographed using silica gel flash chromatography (hexane,hexane/EtOAc (1:1), EtOAc, EtOAc/MeOH (1:1), MeOH, MeOH/HOAc (98:2)).The depsipeptides were found in the EtOAc/MeOH (1:1) fraction. RepeatedHPLC RP18 RP C18 MeCN/H₂O/TFA (55/45/1 30/70/1)—MeCN/H₂O/TFA ((30/70/155/45/1) yielded six the new depsipeptides.

KAHALALDDE FKAHALALIDE F

Final purification was accomplished by HPLC on RP18 RP C18 MeCN/H₂O/TFA(55/45/1). Physical data: [α]D-8° [α]_(D)- 8° (c 4.32, MeOH); ¹H NMR(500 MHz, TFA/DMF); amino acid unit, δ (carbon position, mult, J): Val-14.16 (2, t, J=9.0 Hz), 7.11 (NH on 2, d, J=8.9 Hz), 1.77 (3, m), 0.95(4, m), 0.95 (5, m); Dhb 9.20 (NH on 2, s), 6.48 (3, q, J=6.9 Hz), 1.43(4, d, J=6.6 Hz); Phe 4.68 (2, q, J=6.6 Hz), 8.62 (NH on 2, d, J=6.6Hz), 3.23 (3, dd, J=13.7, 7.2 Hz), 3.00 (3, dd, J=13.7, 9.0 Hz), 7.32(5, d, J=7.2 Hz), 7.28 (6, t, J=7.5 Hz), 7.21 (7, t, J=7.2 Hz); Val-24.36 (2, m), 7.82 (NH on 2, d, J=6.6 Hz), 2.12 (3, m), 0.85 (4, m), 0.77(5, d, J=6.6 Hz); Ileu-1 4.53 (2, m), 8.38 (NH on 2, d, J=9.6 Hz), 1.98(3, m), 0.92 (4, d, J=6.6 Hz), 1.40 (5, m), 1.13 (5, m), 0.88 (6, t,J=7.2 Hz); Thr-1 4.63 (2, t, J=9.3 Hz), 8.12 (NH on 2, d, J=5.7), 5.07(3, dq, J=9.6, 6.0 Hz), 1.18 (4, d, J=6.3 Hz); Ileu-2 4.52 (2, m), 7.72(NH on 2, d, J=8.4 Hz), 1.88 (3, m), 0.88 (4, d, J=6.3 Hz), 1.40 (5, m),1.13 (5, m), 0.88 (6, d t, J=7.2 Hz); Orn 4.48 (2, m), 7.92 (NH on 2, d,J=7.8 Hz), 1.76 (3, m), 1.83 (4, m), 3.10 (5, p, J=5.1 Hz); Pro 4.42 (2,m), 2.12 (3, m), 1.97 ( 3, m), 2.02 (4, m), 1.88 (4, m), 3.75 (5, m),3.68 (5, m); Val-3 4.41 (2, m), 7.90 (NH on 2, d, J=7.2 Hz), 2.12 (3,m), 0.95 (4, m), 0.85 (5, m); Val-4 4.34 (2, m), 7.68 (NH on 2, d, J=8.1Hz), 2.17 (3, m), 0.95 (4, m), 0.90 (5, m); Thr-2 4.46 (2, m), 7.77 (NHon 2, d, J=8.1), 4.21 (3, dq, J=6.3, 3.6 Hz), 1.12 (4, d, J=6.6 Hz);Val-5 4.32 (2, m), 7.85, (NH on 2, d, J=8.1 Hz), 7.82 (NH on (secondconformation) d, J=8.1 Hz), 2.14 (3, m), 0.95 (4, m), 0.90 (5, m);5-MeHex 2.26 (2, m), 1.60 (3, m), 1.20 (4, m), 1.55 (5, m), 0.87 (6, d,J=7.2 Hz), 0.87 (7, d, J=7.2 Hz); 5-MeHex 2.29 (2, m), 1.65 (3, m), 1.40(3, m), 1.13 (4, m), 1.35 (5, m), 0.90 (6, m), 0.90 (7, m) ; ¹³C NMR(125 MHz TFA/DMF): amino acid unit, δ (carbon position); Val-1 70.40170.40 (1), 60.31 (2), 30.75 (3), 19.58 (4), 18.76 (5); Dhb 164.54 (1),130.30 (2), 131.26 (3), 12.68 (4); Phe 171.31 (1), 56.27 (2), 36.79 (3),138.23 (4), 129.86 (5), 128.77 (6), 126.98 (7); Val-2 172-94 172.94 (1),58.57 (2), 32.38 (3), 18.92 (4), 17.60 (5); Ileu-1 171.87 (1), 57.48(2), 38.78 (3), 14.56 (4), 26.78 (5), 11.67 ( 6 ); Thr-1 169.68 (1),57.37 (2), 71.05 (3), 17.34 (4); Ileu-2 171.92 (1), 57.29 (2), 38.01(3), 14.78 (4), 26.55 (5), 11.63 (6); Orn 172.01 (1), 52.87 (2), 29.63(3), 24.39 (4), 40.05 (5); Pro 172.55 (1), 60.23 (2), 29.58 (3), 25.38(4), 48.03 (5); Val-3 171.28 (1), 57.57 (2), 30.54 (3), 19.61 (4), 18.80(5); Val-4 171.83 (1), 59.10 (2), 31.26 (3), 19.45 (4), 18.08 (5); Thr-2170.97 (1), 58.89 (2), 67.36 (3), 19.66 (4); Val-5 172.67 (1), 59.64(2), 30.66 (3), 19.61 (4), 18.43 (5), 5-MeHex 173.83 (1), 36.28 (2),23.99 (3), 38.96 (4), 28.10 (5), 22.54 (6), 22.50 (7); 5-MeHex (secondconformation) 174.08 (1), 33.86 (2), 32.84 (3), 29.75 (4), 34.54 (5),19.51 (6), 11.20 (7) ; IR neat (NaCl): 3287 (s, br), 2964 (s, br), 1646(s), 1528 (s), 1465 (s), 1388 (m), 1228 (m), cm⁻¹; mass spectrum HRFABm/z (fragment, %) 1477.9408 (M⁺+1, 85) (calcd for C₇₅H₁₂₅N₁₄O₁₆:1477.9398); UV (MeOH): λ_(max) 204 (89,630) nm.

Amino acid analysis by GC-MS with a Chirasil-Val column indicates thatKahalalide F consists of 2-D-allo-Ileu, L-Orn, L-Phe, D-Pro, L-Thr,D-Allo-Thr, 3 D-Val and 2 L-Val.

TABLE IITABLE I ¹H and ¹³C NMR Data for Kahalalide F (I)KF in DMF/TFAAmino Acid Carbon ¹³C, ppm^(a) Mult. ¹H, ppm^(b) Multiplicity Valine-1 1170.4 s (NH) 7.11 d, J=8.9 2 60.3 d 4.16 t, J=9.0 3 30.8 d 1.77 m 4 19.6q 0.95 m 5 18.8 q 0.95 m Dehydroamino 1 164.5 s (NH) 9.20 s butyric acid2 130.3 s 3 131.3 d 6.48 q, J=6.9 4 12.7 q 1.43 d, J=6.6 Phenylalanine 1171.3 s (NH) 8.62 d, J=6.6 2 56.3 d 4.68 q, J=6.6 3 36.8 t 3.23 dd,J=13.7, 3.00 7.2 3.00 dd, J=13.7, 9.0 4 138.2 s 5, 5′ 129.9 d 7.32 d,J=7.2 6, 6′ 128.8 d 7.28 t, J=7.5 7 127.0 d 7.21 t, J=7.2 Valine-2 1172.9 s (NH) 7.82 d, J=6.6 2 58.6 d 4.36 m 3 32.4 d 2.12 m 4 18.0 q 0.85m 5 17.6 q 0.77 d, J=6.6 Isoleucine-1 1 171.9 s (NH) 8.38 d, J=0.6 257.5 d 4.53 m 3 38.8 d 1.98 m 4 14.6 q 0.92 d, J=6.6 5 26.8 t 1.40, 1.13m, m 6 11.7 q 0.88 t, J=7.2 Threonine-1 1 169.7 s (NH) 8.12 d, J=5.7 257.4 d 4.63 t, J=9.3 3 71.1 d 5.07 dq, J=9.6, 6.0 4 17.3 q 1.18 d, J=6.3Isoleucine-2 1 171.9 s (NH) 7.72 d, J=8.4 2 57.3 d 4.52 m 3 38.0 d 1.88m 4 14.8 q 0.88 d, J=6.3 5 26.6 t 1.40, 1.13 m, m 6 11.6 q 0.88 t, J=7.2Ornithine 1 172.0 s (NH) 7.92 d, J=7.8 2 52.9 d 4.48 m 3 29.6 t 1.76 m 424.4 t 1.83 m 5 40.1 t 3.10 p, 5.1 Proline 1 172.6 s 2 60.2 d 4.42 m 329.6 t 2.12, 1.97 m, m 4 25.4 t 2.02, 1.88 m, m 5 48.0 t 3.75, 3.68 m, mValine-3 1 171.3 s (NH) 7.90 d, J=7.2 2 57.6 d 4.41 m 3 30.5 d 2.12 m 419.6 q 0.95 m 5 18.8 q 0.85 m Valine-4 1 171.8 s (NH) 7.68 d, J=8.1 259.1 d 4.34 m 3 31.3 d 2.17 m 4 19.5 q 0.95 m 5 18.1 q 0.90 mThreonine-2 1 171.0 s (NH) 7.77 d, J=8.1 2 58.9 d 4.46 m 3 67.4 d 4.21dq, J=6.3, 3.6 4 19.7 q 1.12 d, J=6.6 Valine-5 1 172.7 s (NH) 7.85 d,J=8.1 conf. #2 7.85 d, J=7.81 (NH) 7.82 2 59.6 d 4.32 m 3 30.7 d 2.14 m4 19.6 q 0.95 m 5 18.4 q 0.90 m 5-Methyl- 1 173.8 s Hexanoic acid 2 36.3t 2.26 m 3 24.0 t 1.60 m 4 39.0 t 1.20 m 5 28.1 d 1.55 m 6 22.5 q 0.87d, J=7.2 7 22.5 q 0.87 d, J=7.2 5-Methyl- 1 174.1 s Hexanoic acid 2 33.9t 2.29 m (second 3 32.8 t 1.65, 1.40 m conformation) 4 29.8 t 1.13 m 534.5 d 1.35 m 6 19.5 q 0.90 m 7 11.2 q 0.90 m ^(a)at 125 MHz, DMF signalat 35.2 ppm; ^(b)at 500 MHz, DMF signal at 2.91 ppm.

TABLE ITABLE II In vitro Activity of Kahalalide F KF from Elysiarufescens Assay (M.I.C. μg/mL) Cytoxicity μg/mL (IC50) A-549 2.5 HT-290.25-0.5 Antiviral μg/mL (% reduction) Mv 2 Lu/HSV II 0.5 (95%)CV-1/HSV-1 >8 BHK/VSV >8 Antifungal 6 mm disk 50 μg/disk Aspergillusoryzae 19 mm Penicillium notatum 26 mm Tricophyton mentagrophy 34 mmSaccharomyces cerevisiac neg Candida albicans 16 mm

1. A substially Substantially pure compound Kahalalide F compound, said compound hag having a molecular formula of C₇₅H₁₂₄N₁₄O₁₆, and consisting of five valines, two isoleucines, two threonines, ornithine, dehydroaminobutynic dehydroaminobutyric acid, proline, phenylalanine and 5-methylhexanoic acid; said compound further exhibiting the following physical and chemical properties: [α]_(D)-8° (c 4.32, MeOH); ¹H NMR (500 MHz, TFA/DMF); amino acid unit, δ (carbon position, mult, J): Val-1 4.16 (2, t, J=9.0 Hz), 7.11 (NH on 2, d, J=8,9 8.9 Hz), 1.77 (3, m), 0.95 (4, m), 0.95 (5, m), Dhb 9.20 (NH on 2, s), 6.48 (3, q, J=6.9 Hz), 1.43 (4, d, J=6.6 Hz); Phe 4.68 (2, q, J=6.6 Hz), 8.62 (NH on 2, d, J=6.6 Hz), 3.23 (3, dd, J=13.7, 7.2 Hz), 3.00 (3, dd, J=13.7, 9.0 Hz), 7.32 (5, d, J=7.2 Hz), 7.28 (6, t, J=7.5 Hz), 7.21 (7, t, J=7.2 Hz); Vol-2 Val-2 4.36 (2, m), 7.82 (NH on 2, d, J=6.6 Hz), 2.12 (3, m), 0.85 (4, m), 0.77 (5, d, J=6.6 Hz); Ileu-1 4.53 (2, m), 8.38 (NH on 2, d, J=9.6 Hz), 1.98 (3, m), 0.92 (4, d, J=6.6 Hz), 1.40 (5, m), 1.13 (5, m), 0.88 (6, t, J=7.2 Hz); Thr-1 4.63 (2, t, J=9.3 Hz), 8.12 (HN NH on 2, d, J=5.7), 5.07 (3, dq, J=9.6, 6.0 Hz), 1.18 (4, d, J=6.3 Hz); Ileu-2 4.52 (2, m), 7.72 (NH on 2, d, J=8.4 Hz), 1.88 (3, m), 0.88 (4, d, J=6.3 Hz), 1.40 (5, m), 1.13 (5, m), 0.88 (6, d t, J=7.2 Hz) Orn 4.48 (2, m), 7.92 (NH on 2, d, J=7.8 Hz), 1.76 (3, m), 1.83 (4, m), 3.10 (5, p, J=5.1 Hz); Pro 4.42 (2, m), 2.12 (3, m), 1.97 (3, m), 2.02 (4, m), 1.88 (4, m), 3.75 (5, m), 3.68 (5, m); Val-3 4.41 (2, m), 7.90 (NH on 2, d, J=7.2 Hz), 2.12 (3, m), 0.95 (4, m), 0.85 (5, m); Val-4 4.34 (2, m), 7.68 (NH on 2, d, J=8.1 Hz), 2.17 (3, m), 0.95 (4, m), 0.90 (5, m); Thr-2 4.46 (2, m), 7.77 (NH on 2, d, J=8.1 Hz), 4.21 (3, dq, J=6.3, 3.6 Hz), 1.12 (4, d, J=6.6 Hz); Val-5 4.32 (2, m), 7.85 (HN NH on 2, d, J=8.1 Hz), 7.82 (NH on (second conformation), d, J=8.1 Hz), 2.14 (3, m), 0.95 (4, m), 0.90 (5, m); 5-MeHex 2.26 (2, m), 1.60 (3, m), 1.20 (4, m), 1.55 (5, m), 0.87 (6, d, J=7.2 Hz), 0.87 (7, d, J=72 7.2 Hz); 5-MeHex 2.29 (2, m), 1.65 (3, m), 1.40 (3, m), 1.13 (4, m), 1.35 (5, m), 0.90 (6, m), 0.90 (7, m); ¹³C NMR (125 MHz TFA/DMF): amino acid unit, δ (carbon position); Val-1 170.40 (1), 60.31 (2), 30.75 (3), 19.58 (4), 18.76 (5); Dhb 164.54 (1), 130.30 (2), 131.26 (3), 126.6 12.68 (4); Phe 171.31 (1), 56.27 (2), 36.79 (3), 138.23 (4), 129.86 (5), 128.77 (6), 126.98 (7); Val-2 172.94 (1), 58.57 (2), 32.38 (3), 18.92 (4), 17.60 (5); Ileu-1 171.87 (1), 57.48 (2), 38.78 (3), 14.56 (4), 26.78 (5), 11.67 ( 6 ); Thr-1 169.68 (1), 57.37 (2), 71.05 (3), 17.34 (4); Ileu-2 171.92 (1), 57.29 (2), 38.01 (3), 14.78 (4), 26.55 (5), 11.63 (6); Orn 172.01 (1), 52.87 (2), 29.63 (3), 24.39 (4), 40.05 (5); Pro 172.55 (1), 60.23 (2), 29.58 (3), 25.38 (4) 48.03 (5); Val-3 171.28 (1), 57.57 (2), 30.54 (3), 19.61 (4), 18.80 (5); Val-4 171.83 (1), 59.10 (2), 31.26 (3), 19.45 (4), 18.08 (5); thr-2 Thr-2 170.97 (1), 58.89 (2), 67.36 (3), 19.66 (4); Val-5 172.67 (1), 59.64 (2), 30.66 (3), 19.61 (4), 18.43 (5); 5-MeHex 173.83 (1), 36.28 (2), 23.99 (3), 38.96 (4), 28.10 (5), 22.54 (6), 22.50 (7); 5-MeHex (second conformation) 174.08 (1), 33.86 (2), 32.84 (3), 29.75 (4), 34.54 (5), 19.51 (6), 11.20 (7); IR neat (NaCl): 3287 (s, br), 2964 (s, br), 1646 (s), 1528 (s), 1464 (s), 1388 (m), 1228 (m), cm⁻¹; mass spectrum HRFAB m/z (fragment, %) 1477.9408 (M⁺+1, 85); UV (MeOH): λ_(max) 204 (89,630) nm.
 2. The compound Kahalalide F compound of claim 1, which further has the folowing following non-stereospecific structure:


3. A pharmaceutical compostion composition comprising a pharmaceutical carrier or diluent and the substantially pure compound Kahalalide F, said compoumd compound having a molecular formula of C₇₅H₁₂₄N₁₄O₁₆, and conng consisting of five valines, two isoleucines, two threonines, ornithine, dehydroamiobutyric dehydroaminobutyric acid, proline, phenylalanine and 5-methylhexoic 5-methylhexanoic acid; said compound further exhibiting the following physical and chemical properties: [α]_(D)-8° (c 4.32, MEOH); ¹H NMR (500 MHz, TFA/DMF); amino acid unit, δ (carbon position, mult, J): Val-1 4.16 (2, t, J=9.0 Hz), 7.11 (NH on 2, d, J=8,9 8.9 Hz), 1.77 (3, m), 0.95 (4, m), 0.95 (5, ma m); Dhb 9.20 (NH on 2, s), 6.48 (3, q, J=6.9 Hz), 1.43 (4, d, J=6.6 Hz); Phe 4.68 (2, q, J=6.6 Hz), 8.62 (NH on 2, d, J=6.6 Hz), 3.23 (3, dd, J=13.7, 7.2 Hz), 3.00 (3, dd, J=13.7, 9.0 Hz), 7.32 (5, d, J=7.2 Hz), 7.28 (6, t, J=7.5 Hz), 7.21 (7, t, J=7.2 Hz); Val-2 4.36 (2, m), 7.82 (NH on 2, d, J=6.6 Hz), 2.12 (3, m), 0.85 (4, m), 0.77 (5, d, J=6.6 Hz); Ileu-1 Ileu- 1 4.53 (2, m), 8.38 (NH on 2, d, J=9.6 Hz), 1.98 (3, m), 0.92 (4, d, J=6.6 Hz), 1.40 (5, m), 1.13 (5, m), 0.88 (6, t, J=7.2 Hz); Thr-1 4.63 (2, t, J=9.3 Hz), 8.12 (NH on 2, d, J=5.7), 5.07 (3, dq, J=9.6, 6.0 Hz), 1.18 (4, d, J=6.3 Hz); Ileu-2 4.52 (2, m), 7.72 (NH on 2, d, J=8.4 Hz), 1.88 (3, m), 0.88 (4, d, J=6.3 Hz), 1.40 (5, m), 1.13 (5, m), 0.88 (6, d t, J=7.2 Hz) Orn 4.48 (2, m), 7.92 (NH on 2, d, J=7.8 Hz), 1.76 (3, m), 1.83 (4, m), 3.10 (5, p, J=5.1 Hz); Pro 4.42 (2, m), 2.12 (3, m), 1.97 (3, m), 2.02 (4, m), 1.88 (4, m), 3.75 (5, m), 3.68 (5, m); Val-3 4.41 (2, m), 7.90 (NH on 2, d, J=7.2 Hz), 2.12 (3, m), 0.95 (4, m), 0.85 (5, m); Val-4 4.34 (2, m), 7.68 (NH on 2, d, J=8.1 Hz), 2.17 (3, m), 0.95 (4, m), 0.90 (5, m); Thr-2 4.46 (2, m), 7.77 (NH on 2, d, J=8.1Hz), 4.21 (3, dq, J=6.3, 3.6 Hz), 1.12 (4, d, J=6.6 Hz); Val-5 4.32 (2, m), 7.85, (NH on 2, d, J=8.1 Hz), 7.82 (NH on (second conformation), d, J=8.1 Hz), 2.14 (3, m), 0.95 (4, m), 0.90 (5, m); 5MeHex 2.26 (2, m), 1.60 (3, m), 1.20 (4, m), 1.55 (5, m), 0.87 (6, d, J=7.2 Hz), 0.87 (7, d, J=7.2 Hz); 5-MeHex 2.29 (2, m), 1.65 (3, m), 1.40 (3, m), 1.13 (4, m), 1.35 (5, m), 0.90 (6, m), 0.90 (7, m); ¹³C NMR (125 MHz TFA/DMF): amino acid unit, δ (carbon position); Val-1 170.40 (1), 60.31 (2), 30.75 (3), 19.58 (4), 18.76 (5); Dhb 164.54 (1), 130.30 (2), 131.26 (3), 12.68 (4); Phe 171.31 (1), 56-27 56.27 (2), 36.79 (3), 138.23 (4), 129.86 (5), 128.77 (6), 126.98 (7); Val-2 172.94 (1), 58.27 (2), 32.38 (3), 18.92 (4), 17.60 (5); Ileu-1 171.87 (1), 57.48 (2), 38.78 (3), 14.56 (4), 26.78 (5), 11.67 ( 6 ); Thr-1 169.68 (1), 57.37 (2), 71.05 (3), 17.34 (4); Ileu-2 171.92 (1), 57.29 (2), 38.01 (3), 14.78 (4), 26.55 (5), 11.63 (6); Orn 172.01 (1), 52.87 (2), 29.63 (3), 24.39 (4), 40.05 (5); Pro 172.35 172.55 (1), 60.23 (2), 29.58 (3), 25.38 (4), 48.03 (5); Val-3 171.28 (1), 57.57 (2), 30.54 (3), 19.61 (4), 18.80 (5); Val-4 171.83 (1), 59.10 (2), 31.26 (3), 19.45 (4), 18.08 (5); Thr-2 170.97 (1), 58.89 (2), 67.36 (3), 19.66 (4); Val-5 172.67 (1), 59.64 (2), 30.66 (3), 19.61 (4), 18.43 (5); 5-MeHex 173.83 (1), 36.28 (2), 23.99 (3), 38.96 (4), 28.10 (5), 22.54 (6), 22.50 (7); 5-MeHex (second conformation) 174.08 (1), 33.86 (2), 32.84 (3), 29.75 (4), 34.54 (5), 19.51 (6), 11.20 (7); IR neat (NaCl): 3287 (s, br), 2964 (s, br), 1646 (s), 1528 (s), 1464 (s), 1388 (m), 1228 (m), cm⁻¹; mass spectrum HRFAB m/z (fragment, %) 1477.9408 (M⁺+1, 85); UV (MeOH): λ_(max) 204 (89,630) nm.
 4. The pharmaceutical composition of claim 3, wherein the compound Kahalalide F compound further has the folowing non-streospecific following non-stereospecific structure


5. A method of treating fungal infections in mammals comprising administering to a patient in need of such treatment, an amount of the substantially puxe compound pure Kahalalide F compound or a pharmaceutically acceptable salt thereof, sufficient to slow or stop the growth of the fungal infection; said compound having a molecula molecular formula of C₇₅H₁₂₄N₁₄O₁₆, and consisting of five valines, two isoleucines, two theonines threonines, ornithine, dehydroaminobutiric dehydroaminobutyric acid, proline, phenylalanie phenylalanine and 5-methylhexanoic acid; said compound further exhibiting the folowing following physical and chemical properties: [α]_(D)-8° (c 4.32, MEOH); ¹H NMR (500 MHz, TFA/DMF); amino acid unit, δ (carbon position, mult, J): Val-1 4.16 (2, t, J=9.0 Hz), 7.11 (NH on 2, d, J=8,9 8.9 Hz), 1.77 (3, m), 0.95 (4, m), 0.95 (5, ma m); Dhb 9.20 (NH on 2, s), 6.48 (3, q, J=6.9 Hz), 1.43 (4, d, J=6.6 Hz); Phe 4.68 (2, q, J=6.6 Hz), 8.62 (NH on 2, d, J=6.6 Hz), 3.23 (3, dd, J=13.7, 7.2 Hz), 3.00 (3, dd, J=13.7, 9.0 Hz), 7.32 (5, d, J=7.2 Hz), 7.28 (6, t, J=7.5 Hz), 7.21 (7, t, J=7.2 Hz); Val-2 4.36 (2, m), 7.82 (NH on 2, d, J=6.6 Hz), 2.12 (3, m), 0.85 (4, m), 0.77 (5, d, J=6.6 Hz); Ilue-1 Ileu- 1 4.53 (2, m), 8.38 (NH on 2, d, J=9.6 Hz), 1.98 (3, m), 0.92 (4, d, J=6.6 Hz), 1.40 (5, m), 1.13 (5, m), 0.88 (6, t, J=7.2 Hz); Thr-1 4.63 (2, t, J=9.3 Hz), 8.12 (NH on 2, d, J=5.7), 5.07 (3, dq, J=9.6, 6.0 Hz), 1.18 (4, d, J=6.3 Hz); Ileu-2 4.52 (2, m), 7.72 (NH on 2, d, J=8.4 Hz), 1.88 (3, m), 0.88 (4, d, J=6.3 Hz), 1.40 (5, m), 1.13 (5, m), 0.88 (6, d t, J=7.2 Hz) Orn 4.48 (2, m), 7.92 (NH on 2, d, J=7.8 Hz), 1.76 (3, m), 1.83 (4, m), 3.10 (5, p, J=5.1 Hz); Pro 4.42 (2, m), 2.12 (3, m), 1.97 (3, m), 2.02 (4, m), 1.88 (4, m), 3.75 (5, m), 3.68 (5, m); Val-3 4.41 (2, m), 7.90 (NH on 2, d, J=7.2 Hz), 2.12 (3, m), 0.95 (4, m), 0.85 (5, m); Val-4 4.34 (2, m), 7.68 (NH on 2, d, J=8.1 Hz), 2.17 (3, m), 0.95 (4, m), 0.90 (5, m); Thr-2 4.46 (2, m), 7.77 (NH on 2, d, J=8.1), 4.21 (3, dq, J=6.3, 3.6 Hz), 1.12 (4, d, J=6.6 Hz); Val-5 4.32 (2, m), 7.85, (NH on 2, d, J=8.1 Hz), 7.82 (NH on (second conformation), d, J=8.1 Hz), 2.14 (3, m), 0.95 (4, m), 0.90 (5, m); 5MeHex 2.26 (2, m), 1.60 (3, m), 1.20 (4, m), 1.55 (5, m), 0.87 (6, d, J=7.2 Hz), 0.87 (7, d, J=7.2 Hz); 5MeHex 2.29 (2, m), 1.65 (3, m), 1.40 (3, m) 1.13 (4, m), 1.35 (5, m), 0.90 (6, m), 0.90 (7, m); ¹³C NMR (125 MHz, TFA/DMF): amino acid unit, 6 (carbon position); Val-1 170.40 (1), 60.31 (2), 30.75 (3), 19.58 (4), 18.76 (5); Dhb 164.54 (1), 130.30 (2), 131.26 (3), 12.68 (4); Phe 171.31 (1), 56-27 56.27 (2), 36.79 (3), 138.23 (4), 129.86 (5), 128.77 (6), 126.98 (7); Val-2 172.94 (1), 58.27 (2), 32.38 (3), 18.92 (4), 17.60 (5); Ileu-1 171.87 (1), 57.48 (2), 38.78 (3), 14.56 (4), 26.78 (5), 11.67 ( 6 ); Thr-1 169.68 (1), 57.37 (2), 71.05 (3), 17.34 (4); Ileu-2 171.92 (1), 57.29 (2), 38.01 (3), 14.78 (4), 26.55 (5), 11.63 (6); Orn 172.01 (1), 52.87 (2), 29.63 (3), 24.39 (4), 40.05 (5); Pro 172.35 172.55 (1), 60.23 (2), 29.58 (3), 25.38 (4), 48.03 (5); Val-3 171.28 (1), 57.57 (2), 30.54 (3), 19.61 (4), 18.80 (5); Val-4 171.83 (1), 59.10 (2), 31.26 (3), 19.45 (4), 18.08 (5); Thr-2 170.97 (1), 58.89 (2), 67.36 (3), 19.66 (4); Val-5 172.67 (1), 59.64 (2), 30.66 (3), 19.61 (4), 18.43 (5); 5-MeHex 173.83 (1), 36.28 (2), 23.99 (3), 38.96 (4), 28.10 (5), 22.54 (6), 22.50 (7); 5-MeHex (second conformation) 174.08 (1), 33.86 (2), 32.84 (3), 29.75 (4), 34.54 (5), 19.51 (6), 11.20 (7); IR neat (NaCl): 3287 (s, br), 2964 (s, br), 1646 (s), 1528 (s), 1464 (s), 1388 (m), 1228 (m), cm⁻¹; mass spectrum HRFAB m/z (fragment, %) 1477.9408 (M⁺+1, 85); UV (MeOH): λ_(max) 204 (89,630) nm.
 6. The method of treatment of claim 5, wherein compound Kahalalide F has the following non-stereospecific structure:


7. The method of claim 5, wherein the fungal infection is caused by Aspergillus oryzae.
 8. The method of claim 5, wherein the fungal infection is caused by Penicillium notatum.
 9. The method of claim 5, wherein the fungal infection is caused by Trichophyton mentagrophy.
 10. The method of claim 5, wherein the fungal infection is caused by Candida albicans. 